用于囊性纤维化基因治疗的混合慢病毒载体 rSIV.F/HN 的药理和临床前安全性概况。

IF 16.6 1区 医学 Q1 RESPIRATORY SYSTEM
Alena Moiseenko, Anthony Sinadinos, Ana Sergijenko, Kyriel Pineault, Aarash Saleh, Konradin Nekola, Nathalie Strang, Anastasia Eleftheraki, A Christopher Boyd, Jane C Davies, Deborah R Gill, Stephen C Hyde, Gerry McLachlan, Tim Rath, Michael Rothe, Axel Schambach, Silke Hobbie, Michael Schuler, Udo Maier, Matthew J Thomas, Detlev Mennerich, Manfred Schmidt, Uta Griesenbach, Eric W F W Alton, Sebastian Kreuz
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引用次数: 0

摘要

理由和目标:囊性纤维化(CF)是由 CF 跨膜传导调节器(CFTR)基因突变引起的。CFTR 调节剂可明显改善病情,但约有 10% 的患者仍无反应或不能耐受。本研究分析了rSIV.F/HN--一种优化用于肺部递送的慢病毒载体,包括CFTR蛋白表达、CFTR缺陷的功能校正和基因组整合位点分析,为首次人体临床试验做准备:方法:使用来自 CF 患者(F508del/F508del)的原代人支气管上皮细胞(HBEC)的气液界面培养物以及模拟 I 类(CFTR 空)同基因突变的 CFTR 缺陷永生化人肺上皮细胞系来评估转导效率。定量方法包括一种新颖的近接测定法(PLA),用于检测 CFTR 蛋白的表达。为评估 CFTR 通道活性,进行了乌星室研究。结果:rSIV.F/HN表达了CFTR,并在原代F508del/F508del HBECs和I类细胞中将CFTR介导的氯电流恢复到生理水平。相比之下,小分子 CFTR 调节剂无法实现后者,这凸显了基因疗法治疗这类突变的潜力。rSIV.F/HN-CFTR与增效剂ivacaftor的结合显示出大于相加的效果。基因组整合模式没有显示出位点优势(出现频率≤10%),体外永生化试验中观察到插入突变的风险较低:结论:研究结果表明,rSIV.F/HN是一种很有前景的CF基因治疗载体,它提供了一种可诊断基因突变的治疗方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Pharmacological and pre-clinical safety profile of rSIV.F/HN, a hybrid lentiviral vector for cystic fibrosis gene therapy.

Rationale and objective: Cystic fibrosis (CF) is caused by mutations in the CF Transmembrane Conductance Regulator (CFTR) gene. CFTR modulators offer significant improvements, but approximately 10% of patients remain nonresponsive or are intolerant. This study provides an analysis of rSIV.F/HN, a lentiviral vector optimized for lung delivery, including CFTR protein expression, functional correction of CFTR defects and genomic integration site analysis in preparation for a first-in-human clinical trial.

Methods: Air-liquid interface cultures of primary human bronchial epithelial cells (HBEC) from CF patients (F508del/F508del), as well as a CFTR-deficient immortalized human lung epithelial cell line mimicking Class I (CFTR-null) homozygous mutations, were used to assess transduction efficiency. Quantification methods included a novel proximity ligation assay (PLA) for CFTR protein expression. For assessment of CFTR channel activity, Ussing chamber studies were conducted. The safety profile was assessed using integration site analysis and in vitro insertional mutagenesis studies.

Results: rSIV.F/HN expressed CFTR and restored CFTR-mediated chloride currents to physiological levels in primary F508del/F508del HBECs as well as in a Class I cells. In contrast, the latter could not be achieved by small-molecule CFTR modulators, underscoring the potential of gene therapy for this mutation class. Combination of rSIV.F/HN-CFTR with the potentiator ivacaftor showed a greater than additive effect. The genomic integration pattern showed no site predominance (frequency of occurrence ≤10%), and a low risk of insertional mutagenesis was observed in an in vitro immortalization assay.

Conclusions: The results underscore rSIV.F/HN as a promising gene therapy vector for CF, providing a mutation-agnostic treatment option.

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来源期刊
European Respiratory Journal
European Respiratory Journal 医学-呼吸系统
CiteScore
27.50
自引率
3.30%
发文量
345
审稿时长
2-4 weeks
期刊介绍: The European Respiratory Journal (ERJ) is the flagship journal of the European Respiratory Society. It has a current impact factor of 24.9. The journal covers various aspects of adult and paediatric respiratory medicine, including cell biology, epidemiology, immunology, oncology, pathophysiology, imaging, occupational medicine, intensive care, sleep medicine, and thoracic surgery. In addition to original research material, the ERJ publishes editorial commentaries, reviews, short research letters, and correspondence to the editor. The articles are published continuously and collected into 12 monthly issues in two volumes per year.
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